Reaction products of 3-aminomethyl-3,5,5-trimethylcyclohexylamine and m-phenylenebis(methylamine) with 2,2'-[(1-methylethylidene)bis(4,1-phenyleneoxymethylene)]bisoxirane

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

from 2008-05-07 to 2008-06-20

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Version / remarks:

April 2002

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)

Version / remarks:

2004/73/EC

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

4,4'-Isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane, reaction products with 3-aminomethyl-3,5,5-trimethylcyclohexylamine

EC Number:

500-101-4

EC Name:

4,4'-Isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane, reaction products with 3-aminomethyl-3,5,5-trimethylcyclohexylamine

Cas Number:

38294-64-3

Molecular formula:

C41H68N4O4

IUPAC Name:

Reaction product of 3-aminomethyl-3,5,5-trimethylcyclohexanamine with oligomerisation products of 4,4'-propane-2,2-diyldiphenol with 2-(chloromethyl)oxirane

Details on test material:

NA

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Hungarian Supplier: WOBE Kereskedelmi Kft. H-1164, Budapest, Garmada u. 10.
- Age at study initiation: Young adult, 9 - 10 weeks old, age-matched within one week
- Weight at study initiation: The weight variation in animals involved in the study did not exceed ± 20 % of the mean weight.
- Housing: Individual caging Type II. polypropylene/ polycarbonate
- Diet: Animals received ssniff® SM R/M-Z+H "Autoclavable complete feed for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany ad libitum
- Water: Tap water from municipal supply, as for human consumption from 500 mL bottle ad libitum
- Acclimation period: 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes: 8 - 12 air exchange/hour by central air-condition system
- Photoperiod: 12 hours daily, from 6.00 a.m. to 6.00 p.m.

Study design: in vivo (LLNA)

Vehicle:

propylene glycol

Concentration:

25, 10 , 5, 2.5 % (w/v)

No. of animals per dose:

4 animals/treatment group

Details on study design:

In the main assay, twenty female CBA/Ca mice were allocated to five groups of four animals each:
- Four groups received the appropriate formulation of Reaction products of IPDA with bisphenol A diglycidylether (BADGE) at concentrations of 25, 10, 5 and 2.5 % (w/v)
- the negative control group received the solvent propylene glycol (PG)
Each substance was applied on the external surface of each ear (25 µL/ear) for three consecutive days (Day 0, 1 and 2) at the appropriate concentrations. On Day 5, the cell proliferation in the local lymph nodes was measured by in corporation of tritiated methyl-thymidine (3HTdR) and the values obtained were used to calculate the stimulation indices (SI)

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:

The positive control substance alpha-Hexylcinnamaldehyde (HCA) was examined at a concentration of 25 % in the relevant vehicle. A significant lymphoproliferative response (SI >= 3) was noted for HCA with stimulation index value of 7.5, in accordance with our previous data.

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Value:

5.9

Test group / Remarks:

25 % solution

Results 2

Key result

Parameter:

SI

Value:

6.1

Test group / Remarks:

10 % solution

Results 3

Key result

Parameter:

SI

Value:

5.7

Test group / Remarks:

5 % solution

Results 4

Key result

Parameter:

SI

Value:

4.6

Test group / Remarks:

2.5 % solution

Applicant's summary and conclusion

Interpretation of results:

Category 1 (skin sensitising) based on GHS criteria

Conclusions:

Under the conditions of the present assay Reaction products of IPDA with bisphenol A diglycidylether (BADGE), dissolved in an appropriate solvent up to the maximum feasible concentration (25 %), was shown to have sensitisation potential (sensitiser) in the Local Lymph Node Assay. The EC3 value was below the lowest tested concentration of 2.5 %.

Executive summary:

A Local Lymph Node Assay according to OECD 429 and Commission Directive 2004/73/EC, B.42 was performed with CBA/Ca Ola Hsd mice. The maximum concentration of Reaction products of IPDA with bisphenol A diglycidylether (BADGE) in an acceptable solvent was established according to the principles of the relevant guidelines. The maximum attainable concentration based on solubility was 50 % in Propylene glycol (PG) after heating to approximately 80 °C, but because of a very high viscosity, application of this formulation on the ears of animals would not be possible. The next concentration recommended by the relevant guideline is 25 % and this formulation was considered to be physically acceptable for topical application on the ears. A preliminary irritation/toxicity test indicated that the maximum concentration of test item of 25 % in PG was acceptable.

The cutaneous effect observed in the 25 % dose group was considered not to be a classical irritation response and there were no cutaneous effects in lower doses, where a clear lymph node proliferation effect was observed. Hence, in the absence of confounding effects of significant local or systemic toxicity at the applied concentrations, the proliferation values obtained are considered to reflect the real potential of the test item to cause lymphoproliferation in the Local Lymph Node Assay.

A significant lymphoproliferative response (SI >= 3) was noted for Reaction products of IPDA with bisphenol A diglycidylether (BADGE) at all of the applied concentrations. The proliferation values obtained corresponded to the conventional biological dose-response. With proliferation values above 3 obtained at all of the applied concentration and the clear dose-response observed Reaction products of IPDA with bisphenol A diglycidylether (BADGE) was considered a skin sensitiser.